Crystallization and preliminary crystallographic studies of putative threonyl-tRNA synthetases from Aeropyrum pernix and Sulfolobus tokodaii.
Threonyl-tRNA synthetase (ThrRS) plays an essential role in protein synthesis by catalyzing the aminoacylation of tRNA(Thr) and editing misacylation. ThrRS generally contains an N-terminal editing domain, a catalytic domain and an anticodon-binding domain. The sequences of the editing domain in ThrRSs from archaea differ from those in bacteria and eukaryotes. Furthermore, several creanarchaea including Aeropyrum pernix K1 and Sulfolobus tokodaii strain 7 contain two genes encoding either the catalytic or the editing domain of ThrRS. To reveal the structural basis for this evolutionary divergence, the two types of ThrRS from the crenarchaea A. pernix and S. tokodaii have been overexpressed in Eschericha coli, purified and crystallized by the hanging-drop vapour-diffusion method. Diffraction data were collected and the structure of a selenomethionine-labelled A. pernix type-1 ThrRS crystal has been solved using the MAD method.
Shimizu
Satoru
S
Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuda, Midori-ku, Yokohama 226-8501, Japan.
Juan
Ella Czarina Magat
EC
Miyashita
Yu Ichiro
YI
Sato
Yoshiteru
Y
Hoque
Md Mominul
MM
Suzuki
Kaoru
K
Yogiashi
Masataka
M
Tsunoda
Masaru
M
Dock-Bregeon
Anne Catherine
AC
Moras
Dino
D
Sekiguchi
Takeshi
T
Takénaka
Akio
A
D041522:Aeropyrum / Q000201:enzymology*; D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D008958:Models, Molecular; D011487:Protein Conformation; D013045:Species Specificity; D016944:Sulfolobus / Q000201:enzymology*; D013914:Threonine-tRNA Ligase / Q000737:chemistry* / Q000302:isolation & purification
D019843:Archaeal Proteins; D013914:Threonine-tRNA Ligase